Casein is the main protein found in milk and is present at a concentration of about 3-5 gm / 100 ml. It is actually a heterogeneous mixture of phosphorus containing proteins and not a single compound.
Most protein show a minimum solubility at their iso electric point and this principle is used to isolate casein by adjusting the PH of the milk to 4.6 its isoelectric PH. Casein is also insoluble is ethanol and this property is used to remove unwanted fat from preparation.
Take Milk, 1% acetic acid, ethanol, ether, narrow range PH paper.
Procedure: – Transfer 25 ml of given milk in 250 ml beaker. Add 25 ml distilled water in it. Add 1% acetic acid solution, carefully from sides of beaker, till PH of solution is adjusted to 4.6 (check with PH paper). Digest solution for half an hour on water bath. Decant the super natunt. Wash the ppt in the beaker (10-12 times) with boiling water, decanting the super natunt each time. Transfer the ppt. To a small beaker and cover it with. 95% alcohol (15 ml) and stir vigorously for 10- 15 mins. Filter ppt. through watman filter paper no.1 and press the rpt.within the fold to remove alcohol. Pipets extraction with alcohol till casein obtained is finely, divided and free from fat. Dry ppt of carse in an ovea at 110 C.
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